Evaluation of a polymerase chain reaction method for the detection of enteroviruses in groundwater impacted by reclaimed wastewater
AuthorKatner, Adrienne Lee.
Committee ChairGerba, Charles P.
MetadataShow full item record
PublisherThe University of Arizona.
RightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
AbstractThe primary objective of this study was to evaluate the applicability of polymerase chain reaction (PCR) for the detection of enteroviruses in groundwater impacted by reclaimed wastewater. Samples were analyzed by conventional cell culture assay and the reverse transcriptase polymerase chain reaction (RT-PCR), semi-nested PCR, sequencing and dot blot probe hybridization. Viruses were detected in 4/20 samples by cell culture compared to 9/20 by RT-PCR. Semi-nested PCR and probe hybridization increased detection sensitivity to 11/20 and 12/20, respectively. Sequencing of seminested PCR products confirmed enteroviral origin of the amplified sequences, suggesting no cross-reactivity of the primers. Chlorine inactivated viruses were not concentrated as efficiently as cell culture detectable virus (90 percent less efficient). Their ability to be detected by PCR decreased by 90% after five days in secondary effluent and nine days in tap water. These results suggest that PCR is a more sensitive approach for the detection of enteroviruses in the environment. Also, chlorine inactivated viruses are less likely to be detected by PCR than cell culture infectious viruses.
Degree ProgramSoil, Water and Environmental Science